WO2025150943A1 - Novel compound and uses thereof - Google Patents

Abstract

The present invention provides a novel compound and a pharmaceutical composition comprising same for preventing or treating diseases associated with a reduction in Sox9 expression. Accordingly, the diseases associated with a reduction in Sox9 expression can be effectively prevented or treated.

Description

Novel compounds and their uses

The present invention relates to a novel compound, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof, and a use thereof for preventing or treating diseases associated with decreased expression of SOX9.

Osteoarthritis (OA), also known as degenerative arthritis, is a disease that currently affects more than 7% of the world's population (approximately 528 million people). The general treatment for osteoarthritis mainly consists of artificial joint replacement surgery or pain relief treatment through the prescription of cartilage protectors such as hyaluronic acid and anti-inflammatory drugs. However, due to the aging population, the incidence of osteoarthritis is increasing worldwide, and accordingly, the development of direct osteoarthritis treatments rather than simple pain relief is required. One of the causes of osteoarthritis is the gradual loss of cartilage resulting from the destruction of articular cartilage tissue due to the dysregulation of enzymes in the anabolic and catabolic pathways of articular cartilage proteins. Therefore, research is being conducted to inhibit the progression of osteoarthritis by developing disease-modifying OA drugs (DMOADs) that target these regulatory factors. The recently developed Lorecivivint (SM04690) is an example of an osteoarthritis DMOAD.

SOX9 (SRY-Box Transcription Factor 9), a transcription factor that is one of the atypical proteins that plays a key role in regulating the expression of extracellular matrix genes in cartilage cells, is known to regulate the expression of cartilaginous extracellular matrix genes such as collagen required for bone joints. In particular, it was confirmed that the expression and transcriptional activity of SOX9 were reduced in the osteoarthritis patient group, and it was observed that the expression levels of SOX9 and target proteins were significantly lowered depending on the severity of the patient. One of the mechanisms that regulates the activity of transcription factors is the aggregation of transcription factors. As the aggregation phenomenon is induced, the transcription factor aggregates can regulate the expression of genes. That is, when the aggregation phenomenon of the SOX9 transcription factor is induced, the expression and transcriptional effect of SOX9 can be increased through the aggregates.

Therefore, research and development of DMOADs that can directly treat osteoarthritis by inducing aggregation of SOX9 transcription factors and increasing SOX9 transcription efficiency are required.

One object of the present invention is to provide a novel compound capable of inducing the aggregation phenomenon of SOX9 transcription factor, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof.

One object of the present invention is to provide a pharmaceutical composition for preventing or treating a disease associated with decreased expression of SOX9 by using a novel compound capable of inducing aggregation of SOX9 transcription factor, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof.

Each description and embodiment disclosed in this specification may also be applied to each other description and embodiment. That is, all combinations of various elements disclosed in this specification fall within the scope of the present invention. In addition, the scope of the present invention is not limited by the specific description described below.

One aspect of the present invention provides a compound represented by the following chemical formula 1, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

In the above chemical formula 1, one of Y ¹ and Y ² is N, and the other is S, O, or NR ^a1 .

In the above chemical formula 1, X ¹ and X ² are each independently O, S or NR ^a2 .

In the above chemical formula 1, U is NR ⁿ² , a 5- to 7-membered heterocyclyl containing 1 to 3 heteroatoms N, or these linked to each other.

In the above chemical formula 1, Z ¹ is C _1-6 alkylene.

In the above chemical formula 1, Z ² is a direct bond, C _1-6 alkylene, -NR ⁿ³ CO-, or a 5- to 7-membered heteroaryl containing 1 to 3 heteroatoms selected from N, O, and S.

In the above chemical formula 1, R ^a1 and R ^a2 are each independently H or C _1-6 alkyl.

In the above chemical formula 1, R ⁿ¹ , R ⁿ² and R ⁿ³ are each independently H or C _1-6 alkyl.

In the above chemical formula 1, when Z ² is C _1-6 alkylene, any carbon of the alkylene is optionally substituted with halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, carboxy, C _1-6 alkoxycarbonyl, carbamoyl, C _1-6 alkylcarbamoyl, di(C _1-6 alkyl)carbamoyl, cyano, nitro, oxo or C _6-12 aryl.

In the above chemical formula 1, Z ³ is a direct bond or -C(=O)-.

In the above chemical formula 1, ring A is a fused benzoheterocyclyl in which a benzene ring is fused to a 6- to 14-membered aryl, a partially unsaturated 9- to 14-membered bicyclic carbocyclyl, a 5- to 12-membered heteroaryl comprising 1 to 3 heteroatoms selected from O, N or S, a 5- to 8-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S, or a 5- to 7-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S.

In the above chemical formula 1, ring A is optionally substituted with 1 to 3 R ^A .

R ^A is selected from the group consisting of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, cyano, nitro, and oxo.

In the above chemical formula 1, ring E is a 6- to 14-membered aryl, a partially unsaturated 9- to 14-membered bicyclic carbocyclyl, a 5- to 12-membered heteroaryl comprising 1 to 3 heteroatoms selected from O, N or S, a 5- to 12-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S, or a fused heterocycloaryl in which a 5- to 7-membered heterocyclic ring comprising 1 to 3 heteroatoms selected from O, N or S is fused to phenyl.

In the above chemical formula 1, ring E is optionally substituted with 1 to 3 R ^E .

R ^E is selected from the group consisting of halo; hydroxy; C _1-6 alkoxy; amino; C _1-6 alkylamino; di(C _1-6 alkyl)amino; carboxy; C _1-6 alkoxycarbonyl; carbamoyl; C _1-6 alkylcarbamoyl; di(C _1-6 alkyl)carbamoyl; halosulfonyl; sulfoxy; C _1-6 alkylsulfonyl; cyano; nitro; oxo; or C _1-6 alkyl optionally substituted with one or more of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, nitro, and cyano.

In the above chemical formula 1, one of Y ¹ and Y ² can be N, and the other can be S, O, or NR ^a1 . For example, Y ¹ can be N, and Y ² can be S, O, or NR ^a1 , or Y ² can be N, and Y ¹ can be S, O, or NR ^a1 . In some implementations, one of Y ¹ and Y ² can be N, and the other can be S or O. For example, Y ¹ can be N, and Y ² can be S, or Y ¹ can be S or O, and Y ² can be N.

In the above chemical formula 1, X ¹ and X ² can each independently be O, S or NR ^a2 . X ¹ and X ² can be the same or different. For example, X ¹ and X ² can each be O.

In the above formula 1, U can be NR ⁿ² , a 5- to 7-membered heterocyclyl comprising 1 to 3 heteroatoms N, or a combination thereof linked to each other. In some embodiments, U can be NR ⁿ² , a 5- or 6-membered heterocyclyl comprising 1 to 2 heteroatoms N, or a combination thereof linked to each other.

In some implementations, U can be selected from NR ⁿ² ; piperidinediyl; piperazinediyl; or piperidinediyl or piperazinediyl wherein NR ²ⁿ are linked to each other.

In some embodiments, U can be selected from the group consisting of NR ⁿ² and the following structures: In the following structures, * ¹ is connected to a carbon atom of -C(=X ² )- in Chemical Formula 1, and * ² can be connected to Z ² .

The above R ^a1 and R ^a2 can each independently be H or C _1-6 alkyl. In some embodiments, R ^a1 and R ^a2 can each independently be H or C _1-4 alkyl. For example, R ^a1 and R ^a2 can each independently be H or methyl.

In the above formula 1, R ⁿ¹ , R ⁿ² , and R ⁿ³ can each independently be H or C _1-6 alkyl. In some embodiments, R ⁿ¹ , R ⁿ² , and R ⁿ³ can each independently be H or C _1-4 alkyl. For example, R ⁿ¹ , R ⁿ² , and R ⁿ³ can each independently be H or methyl.

In the above formula 1, Z ¹ can be C _1-6 alkylene. In some embodiments, Z ¹ can be C _1-4 alkylene. In some embodiments, Z ¹ can be straight-chain alkylene. In some embodiments, Z ¹ can be methylene, ethylene, propylene, butylene, pentylene, or hexylene.

In the above formula 1, Z ² can be a direct bond, C _1-6 alkylene, -NR ⁿ³ CO- or a 5- to 7-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O and S. The -NR ⁿ³ CO- can be * ³ -NR ⁿ³ CO-* ⁴ , wherein * ³ can be connected to U in the formula 1, and * ⁴ can be connected to ring E. In some embodiments, Z ² can be a direct bond, C _1-4 alkylene, -NR ⁿ³ CO- or a 5- or 6-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O and S. For example, Z ² can be a direct bond, methylene, ethylene, propylene, butylene, -NHCO- or dithiazodiyl.

In the above formula 1, when Z ² is C _1-6 alkylene, any carbon of the alkylene can be optionally substituted with halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, carboxy, C _1-6 alkoxycarbonyl, carbamoyl, C _1-6 alkylcarbamoyl, di(C _1-6 alkyl)carbamoyl, cyano, nitro, oxo or C _6-12 aryl. In some embodiments, any carbon of the alkylene can be optionally substituted with carboxy, C _1-6 alkoxycarbonyl or C _6-12 aryl. For example, any carbon of the alkylene can be substituted with methoxycarbonyl or phenyl. The carbon atom adjacent to ring E in the alkylene of Z ² can be substituted.

In some embodiments, when U is a 5-7 membered heterocyclyl comprising 1 to 3 heteroatoms N or NR ⁿ² and a 5-7 membered heterocyclyl comprising 1 to 3 heteroatoms N linked to each other, Z ² can be a direct bond or C _1-6 alkylene.

In the above chemical formula 1, Z ³ can be a direct bond or -C(=O)-.

In some embodiments, the compound represented by Chemical Formula 1 may be selected from a compound represented by Chemical Formula 2 or Chemical Formula 3 below.

[Chemical formula 2]

[Chemical Formula 3]

In the above chemical formulas 2 and 3, Z ¹ , Z ² , X ¹ , X ² , U, R ⁿ¹ , R ⁿ² , ring A and ring E are as defined in the above chemical formula 1.

In some implementations, Y ¹ of formula 2 and Y ² of formula 3 can each independently be S or O.

In the above chemical formula 1, ring A can be a fused benzoheterocyclyl in which a benzene ring is fused to a 6- to 14-membered aryl, a partially unsaturated 9- to 14-membered bicyclic carbocyclyl, a 5- to 12-membered heteroaryl comprising 1 to 3 heteroatoms selected from O, N or S, a 5- to 8-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S, or a 5- to 7-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S. In some embodiments, ring A can be a fused benzoheterocyclyl wherein a benzene ring is fused to a 6- to 12-membered aryl, a partially unsaturated 9- or 10-membered bicyclic carbocyclyl, a 6- to 10-membered heteroaryl comprising 1 or 2 heteroatoms selected from O, N or S, a 5- to 7-membered heterocyclyl comprising 1 or 2 heteroatoms selected from O, N or S, or a 5- to 7-membered heterocyclyl comprising 1 or 2 heteroatoms selected from O, N or S.

In some embodiments, ring A can be phenyl, biphenylyl, tetrahydronaphthyl, pyranyl, pyranonyl(oxopyranyl), benzopyranyl, or benzopyronyl. The benzopyronyl can be coumarinyl.

In some implementations, ring A is , or and wherein the ring A can be optionally substituted with 1 to 3 R ^A .

In some embodiments, Z ³ is a direct bond and ring A is , or Either; or

Z ³ is -C(=O)-, and ring A is or It could be.

In the above formula 1, ring A can be optionally substituted with 1 to 3 R ^A . R ^A can be selected from the group consisting of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, cyano, nitro, and oxo. In some embodiments, R ^A can be selected from the group consisting of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, and oxo.

In some implementations, R ^A can be selected from the group consisting of halo, hydroxy, C _1-4 alkoxy, amino, C _1-4 alkylamino, di(C _1-4 alkyl)amino, C _1-4 alkyl, cyano, nitro, and oxo.

In some implementations, R ^A can be selected from the group consisting of F, Cl, Br, hydroxy, methoxy, dimethylamino, methyl, or oxo.

In some embodiments, ring A can be selected from the following chemical structures:

.

In the above chemical formula 1, ring E can be a 6- to 14-membered aryl, a partially unsaturated 9- to 14-membered bicyclic carbocyclyl, a 5- to 12-membered heteroaryl comprising 1 to 3 heteroatoms selected from O, N or S, a 5- to 12-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S, or a fused heterocycloaryl in which a 5- to 7-membered heterocyclic ring comprising 1 to 3 heteroatoms selected from O, N or S is fused to phenyl. In some embodiments, ring E can be a 6-12 membered aryl, a partially unsaturated 9-12 membered bicyclic carbocyclyl, a 5-12 membered heteroaryl comprising 1 or 2 heteroatoms selected from O, N or S, a 5-8 membered heterocyclyl comprising 1 or 2 heteroatoms selected from O, N or S, or a fused heterocycloaryl wherein phenyl is fused to a 5-7 membered heterocyclic ring comprising 1 or 2 heteroatoms selected from O, N or S. In some embodiments, ring E can be a 6-10 membered aryl, a partially unsaturated 9- or 10-membered bicyclic carbocyclyl, a 5-10 membered heteroaryl comprising 1 or 2 heteroatoms selected from O, N or S, a 5-7 membered heterocyclyl comprising 1 or 2 heteroatoms selected from O, N or S, or a fused heterocycloaryl wherein phenyl is fused to a 5- or 6-membered heterocyclic ring comprising 1 or 2 heteroatoms selected from O, N or S.

In some embodiments, ring E can be phenyl, naphthalenyl(naphthyl), indanyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, imidazolyl, pyrazolyl, triazolyl, thiophenyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, morpholinyl, thiomorpholinyl, piperidinyl, hexahydroxypyridazinyl, hexahydroxypyrimidinyl, piperazinyl, indolyl, indazolyl, benzoimidazolyl, quinolinyl, isoquinolinyl, cinolinyl, quinazolinyl, quinoxalinyl, phthalazinyl, or benzodioxolyl. For example, ring E can be phenyl, naphthalenyl(naphthyl), pyridinyl, pyridazinyl, pyrimidinyl, imidazolyl, thiophenyl, morpholinyl, thiomorpholinyl, piperazinyl, indolyl, quinolinyl or benzodioxolyl.

In some embodiments, ring E is selected from the following chemical structures; ring E can be optionally substituted with 1 to 3 R ^E .

.

In the above chemical formula 1, ring E may be optionally substituted with 1 to 3 R ^E . When R ^E is 2 or more, each R ^E may be the same or different.

R ^E can be selected from the group consisting of halo; hydroxy; C _1-6 alkoxy; amino; C _1-6 alkylamino; di(C _1-6 alkyl)amino; carboxy; C _1-6 alkoxycarbonyl; carbamoyl; C _1-6 alkylcarbamoyl; di(C _1-6 alkyl)carbamoyl; halosulfonyl; sulfoxy; C _1-6 alkylsulfonyl; cyano; nitro; oxo; or C _1-6 alkyl optionally substituted with one or more of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, nitro, and cyano. In some embodiments, R ^E is halo; hydroxy; C _1-4 alkoxy; amino; C _1-4 alkylamino; di(C _1-4 alkyl)amino; carboxy; C _1-4 alkoxycarbonyl; carbamoyl; C _1-4 alkylcarbamoyl; di(C _1-4 alkyl)carbamoyl; halosulfonyl; sulfoxy; C _1-6 alkylsulfonyl; cyano; nitro; oxo; or C 1-4 alkyl optionally substituted with one or more of halo, hydroxy, C _1-4 alkoxy, amino, C _1-4 alkylamino, di(C _1-4 alkyl ₎ amino, nitro, and cyano.

In some embodiments, R ^E can be selected from the group consisting of halo; hydroxy; C _1-6 alkoxy; amino; C _1-6 alkylamino; di(C _1-6 alkyl)amino; halosulfonyl; sulfoxy; C _1-6 alkylsulfonyl; cyano; nitro; oxo; C _1-6 alkyl, C 1-6 haloalkyl, _or C _1-6 hydroxyalkyl. In some embodiments, R ^E can be selected from the group consisting of halo; hydroxy; C _1-4 alkoxy; amino; C _1-4 alkylamino; di(C _1-4 alkyl)amino; halosulfonyl; sulfoxy; C _1-4 alkylsulfonyl; cyano; nitro; oxo; C _1-4 alkyl, C _1-4 haloalkyl, or C _1-4 hydroxyalkyl.

In some implementations, R ^E can be selected from the group consisting of methoxy, hydroxy, amino, F, Cl, Br, trifluoromethyl, cyano, hydroxymethyl, methyl, fluorosulfonyl, nitro and oxo.

In some embodiments, ring E can be selected from the following chemical structures:

.

In some embodiments, the compound represented by the above formula 1 can be represented by the following formulae I to III.

[Chemical Formula I]

In the above formula I, Y ¹ is S, O or NR ^a1 ; R ^a1 is H or C _1-6 alkyl; R ⁿ¹ and R ⁿ² are each independently H or C _1-6 alkyl; Z ⁴ is a direct bond or C _1-4 alkylene; R ¹ can be H, halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, carboxy, C _1-6 alkoxycarbonyl, carbamoyl, C _1-6 alkylcarbamoyl, di(C _1-6 alkyl)carbamoyl, cyano, nitro, oxo or C _6-12 aryl.

In the above formula I, ring A is a fused benzoheterocyclyl in which a benzene ring is fused to a 6- to 14-membered aryl, a partially unsaturated 9- to 14-membered bicyclic carbocyclyl, a 6- to 12-membered heteroaryl comprising 1 to 3 heteroatoms selected from O, N or S, a 5- to 8-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S, or a 5- to 7-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S; ring A is optionally substituted with 1 to 3 R ^A ; R ^A can be selected from the group consisting of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, cyano, nitro and oxo.

In the above formula I, ring E is a 6-14 membered aryl, a partially unsaturated 9-14 membered bicyclic carbocyclyl, a 5-12 membered heteroaryl comprising 1 to 3 heteroatoms selected from O, N or S, a 5-12 membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S, or a fused heterocycloaryl in which a 5-7 membered heterocyclic ring comprising 1 to 3 heteroatoms selected from O, N or S is fused to phenyl; ring E is optionally substituted with 1 to 3 R ^E ; R ^E is halo; hydroxy; C _1-6 alkoxy; amino; C _1-6 alkylamino; di(C _1-6 alkyl)amino; carboxy; C _1-6 alkoxycarbonyl; carbamoyl; C _1-6 alkylcarbamoyl; di(C _1-6 alkyl)carbamoyl; halosulfonyl; sulfoxy; C _1-6 alkylsulfonyl; cyano; nitro; oxo; or C 1-6 alkyl optionally substituted with one or more of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, nitro, and cyano _.

[Chemical Formula II]

In the above chemical formula II, Y ² is S, O or NR ^a1 ; R ^a1 is H or C _1-6 alkyl; R ⁿ¹ and R ⁿ² are each independently H or C _1-6 alkyl; Z ⁴ is a direct bond or C _1-4 alkylene; R ¹ can be H, halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, carboxy, C _1-6 alkoxycarbonyl, carbamoyl, C _1-6 alkylcarbamoyl, di(C _1-6 alkyl)carbamoyl, cyano, nitro, oxo or C _6-12 aryl.

In the above formula II, ring A ¹ is a 6- to 14-membered aryl or a partially unsaturated 9- to 14-membered bicyclic carbocyclyl; ring A ¹ is optionally substituted with 1 to 3 R ^A ; R ^A can be selected from the group consisting of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, cyano, nitro and oxo. In some embodiments, ring A ¹ can be a 6- to 14-membered aryl.

In the above formula II, ring E is a 6- to 14-membered aryl, a 5- to 12-membered heteroaryl comprising 1 to 3 heteroatoms selected from O, N or S, a 5- to 12-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S, or a fused heterocycloaryl in which a 5- to 7-membered heterocyclic ring comprising 1 to 3 heteroatoms selected from O, N or S is fused to phenyl; ring E is optionally substituted with 1 to 3 R ^E ; R ^E can be selected from the group consisting of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, carboxy, C _1-6 alkoxycarbonyl, carbamoyl, C _1-6 alkylcarbamoyl, di(C _1-6 alkyl)carbamoyl, halosulfonyl, sulfoxy, C _1-6 alkylsulfonyl, cyano, nitro and oxo.

In some embodiments, the compound represented by Formula 1 can be selected from a compound represented by Formula IA, Formula IB, Formula IC, Formula ID, Formula IE, or Formula IIA.

[Chemical Formula III]

In the above formula III, U ¹ is a direct bond or NR ⁿ² ; at least one of Y ³ and Y ⁴ is N, and the others are CH; Z ⁵ is a direct bond, C _1-6 alkylene, -NR ⁿ³ CO-, or a 5- to 7-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O and S; R ⁿ¹ , R ⁿ² and R ⁿ³ are each independently H or C _1-6 alkyl; and s can be 0 or 1.

In the above formula III, ring A can be a fused benzoheterocyclyl in which a benzene ring is fused to a 6- to 14-membered aryl, a partially unsaturated 9- to 14-membered bicyclic carbocyclyl, a 5- to 12-membered heteroaryl comprising 1 to 3 heteroatoms selected from O, N or S, a 5- to 8-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S, or a 5- to 7-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S.

In the above formula III, ring A is optionally substituted with 1 to 3 R ^A ; R ^A can be selected from the group consisting of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, cyano, nitro and oxo.

In the above formula III, ring E can be a 6- to 14-membered aryl, a partially unsaturated 9- to 14-membered bicyclic carbocyclyl, a 5- to 12-membered heteroaryl comprising 1 to 3 heteroatoms selected from O, N or S, a 5- to 12-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S, or a fused heterocycloaryl in which a 5- to 7-membered heterocyclic ring comprising 1 to 3 heteroatoms selected from O, N or S is fused to phenyl.

In the above formula III, ring E is optionally substituted with 1 to 3 R ^E ; R ^E can be selected from the group consisting of halo; hydroxy; C _1-6 alkoxy; amino; C _1-6 alkylamino; di(C _1-6 alkyl)amino; carboxy; C _1-6 alkoxycarbonyl; carbamoyl; C _1-6 alkylcarbamoyl; di(C _1-6 alkyl)carbamoyl; halosulfonyl; sulfoxy; C _1-6 alkylsulfonyl; cyano; nitro; oxo; or C _1-6 alkyl optionally substituted with one or more of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, nitro and cyano.

[Chemical Formula IA]

[Chemical Formula IB]

[Chemical Formula IC]

[chemical formula ID]

[chemical formula IE]

[Chemical formula IIA]

In the above Chemical Formulas IA, IB, IC, ID, IE and IIA, R ^A is selected from the group consisting of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, cyano, nitro and oxo; n, m1, m2, o1, o2, p and q are each independently an integer from 0 to 3; the sum of m1 and m2 and o1 and o2 is each 3 or less; Y ¹ , Y ² , R ⁿ¹ , R ⁿ² , Z ⁴ , R ¹ and ring E are as described in the above Chemical Formulas I and II.

In some embodiments, the compound represented by the formula 1 may be selected from a compound represented by the following formula:

.

In some embodiments, the compound represented by the chemical formula 1 can be synthesized according to the following reaction scheme 1.

[Reaction Formula 1]

In the above reaction scheme 1, Y ¹ , Y ² , Z ³ , ring A, X ¹ , X ² , Z ¹ , Z ² , R ¹ⁿ , U and ring E are as described in the above chemical formula 1.

Specifically, in step 1 of the above reaction scheme 1, ring A is optionally connected via Z ³ , and intermediate a including a 5-membered heteroarene ring substituted with amino can be reacted with cyclic intermediate 1 to synthesize intermediate b. The reaction of step 1 may be carried out under the condition of the presence of a suitable organic solvent that dissolves intermediate a or intermediate i, or may be carried out in a molten state without using an organic solvent at a temperature higher than the melting point (120° C.) of intermediate i. For example, the reaction of step 1 may be performed at about 120° C. to 180° C. for 1 hour to 5 hours.

In step 2 of the above reaction scheme 1, the compound represented by chemical formula 1 can be prepared by reacting intermediate b with amine intermediate c containing ring E. The above step 2 can be performed under anhydrous conditions, and can be performed using an organic solvent such as dioxin at a temperature of about 80° C. to 140° C. for 5 to 24 hours.

The above intermediate a can be synthesized according to the following reaction scheme 2.

[Reaction Formula 2]

In the above reaction scheme 2, rings A and Z ³ are as described in the above chemical formula 1.

Specifically, in step 1 of the above reaction scheme 2, a bromoacetyl compound can be synthesized by reacting an acetyl compound including ring A with an excess of tetrabutylammonium tribromide (CAS: 38932-80-8) in an organic solvent. In the reaction, the organic solvent may be, but is not limited to, CH ₂ Cl ₂ , and the reaction temperature may be about 10° C. to 50° C., for example, room temperature.

In step 2 of the above reaction scheme 2, the obtained product of step 1 and thiourea can be heated in ethanol to obtain intermediate a'. Intermediate a' can be provided as intermediate a of the above reaction scheme 1.

As used herein, the term "halogen" or "halogen atom" refers to an atom belonging to Group 17 of the periodic table. Halogen atoms include F, Cl, Br, I, etc. The term "halo" refers to a halogen substituent.

The term "alkyl" refers to a fully saturated branched or unbranched (or straight-chain or linear) hydrocarbon. The alkyl can be a substituted or unsubstituted alkyl. The C _1-20 alkyl can be, for example, a C _1-15 , C _1-10 , or C _1-6 alkyl. The C _1-6 alkyl can be a C _1-5 , C _1-4 , C _1-3 , or C _1-2 alkyl. The alkyl can be, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, neopentyl, iso-amyl, or n-hexyl.

The term "haloalkyl" refers to alkyl substituted with one or more halo or halogen, and includes dihaloalkyl, trihaloalkyl, and the like. "Haloalkyl" may include perhaloalkyl where all of the hydrogens of the alkyl are replaced with halogen.

The term "hydroxy" refers to the -OH functional group (hydroxyl group).

The term "hydroxyalkyl" refers to alkyl substituted with a hydroxy group.

The term "carbonyl" refers to -C(=O)-.

The term "alkoxy" refers to an alkyl group bonded to an oxygen atom. The C _1-20 alkoxy can be, for example, an alkoxy having C _1-15 , C _1-10 , or C _1-6 . The C _1-6 alkoxy can be an alkoxy having C _1-5 , C _1-4 , C _1-3 , or C _1-2 . The alkoxy can be methoxy, ethoxy, propoxy, butoxy, and the like.

The term "alkoxyalkyl" refers to an alkoxy bonded to alkyl. The C _2-20 alkoxyalkyl can be, for example, a C _2-15 , C _2-10 , or C _2-6 alkoxyalkyl. For example, the C _2-20 alkoxyalkyl can be (C _1-10 alkoxy)-(C _1-10 alkyl), (C _1-6 alkoxy)-(C _1-6 alkyl), etc., and the carbon numbers of the alkoxy group and the alkyl group can be the same or different. The above alkoxy can be, for example, methoxymethyl, methoxyethyl, ethoxymethyl, ethoxyethyl, methoxypropyl, ethoxypropyl, and the like.

The term "amino" refers to -NH ₂ .

The term "amine group" refers to a substituent in which one, two or all three hydrogens of ammonia are replaced by an organic functional group, including primary amines, secondary amines and tertiary amines, and includes an amino group.

The term “alkylamine” refers to an amine in which one of the H atoms in the amino group (-NH ₂ ) is replaced by an alkyl group.

The term “di(alkyl)amine” refers to an amine in which both H of the amino (-NH ₂ ) group are replaced by alkyl. The two alkyl groups in a di(alkyl)amine can be the same or different.

The term "nitro" refers to -NO ₂ .

The term "cyano", -CN, refers to a functional group consisting of a triple bond between a carbon atom and a nitrogen atom.

The term "carboxy" refers to -COOH. A salt of carboxy is the conjugate base of a carboxylic acid.

The term "alkoxycarbonyl" refers to a monovalent substituent in which the -OH of carboxy is replaced by an alkoxy. For example, C _1-6 alkoxycarbonyl refers to -C(=O)- substituted with a C _1-6 alkyl.

The term "carbamoyl" refers to -CONH ₂ .

The term “alkylcarbamoyl” refers to a substituent in which one hydrogen atom of -NH ₂ in carbamoyl is replaced by an alkyl.

The term “dialkylcarbamoyl” refers to a substituent in which the two hydrogen atoms of -NH ₂ in carbamoyl are each replaced by alkyl. The two alkyls in a dialkylcarbamoyl can be the same or different.

The term “sulfonyl” or “sulfonyl” refers to the group -SO ₂ -.

The term "halosulfonyl" refers to a monovalent substituent, which is a sulfonyl substituted with a halogen.

The term "carbocyclyl" refers to a monovalent non-aromatic hydrocarbon ring substituent. Carbocyclyl can be fully saturated or partially unsaturated, and can have a monocyclic structure or a bicyclic or tricyclic structure with fused, bridged or spiro structures. For example, a carbocyclyl is a phenyl having a saturated heterohexane ring fused thereto, including tetrahydronaphthyl, etc. A carbocyclyl can be a substituent having 9 to 14 carbon atoms in the ring.

The term “cycloalkyl” refers to a saturated non-aromatic monocyclic, bicyclic, or tricyclic hydrocarbon group. A cycloalkyl can contain from 3 to 20, for example from 5 to 10, from 3 to 8, or from 3 to 6 carbon atoms. A monocyclic cycloalkyl can be, for example, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. A bicyclic cycloalkyl can be, for example, bornyl, decahydronaphthyl, bicyclo[2.1. 1]hexyl, bicyclo[2.2. 1]heptyl, or bicyclo[2.2. 2]octyl. A tricyclic cycloalkyl can be, for example, adamantyl.

The term “cycloalkane ring” refers to a saturated non-aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring. The cycloalkane ring can be a complete (non-radical) functional group of the cycloalkyl. It can contain 3 to 20 carbon atoms, for example 5 to 10, 3 to 8, or 3 to 6 carbon atoms. The monocyclic cycloalkane ring can be, for example, a cyclopropane ring, a cyclobutane ring, a cyclopentane ring, or a cyclohexane ring.

The term "aryl" refers to an aromatic hydrocarbon ring group. Aryl includes a hydrocarbon ring group having an aromaticity by linking multiple aryls. The C _6-30 aryl can be, for example, a C _6-14 , C _6-12 or C _6-10 aryl. The aryl can have a monocyclic structure, or a bicyclic or tricyclic structure of a fused, bridged or spiro structure. The aryl can be phenyl, naphthyl or biphenyl.

The term "arylalkyl" refers to an alkyl substituted with an aryl.

The term "aryloxy" refers to an aryl bonded to an oxygen atom.

The term "heteroaryl" or "heteroarene" refers to a monocyclic or bicyclic aromatic compound or substituent containing one or more heteroatoms, the remaining ring atoms being carbon. The heteroaryl may include a ring group having aromaticity as a whole by substituting oxo (=O), thioxo (=S), etc. on unsaturated ring atoms, even if the ring itself is partially unsaturated. The heteroaryl may contain, for example, 1 to 5, 1 to 3, 1 or 2 heteroatoms, and may contain 5 to 12 ring members. “Heteroaryl” can be, for example, pyrrolyl, furanyl, thiophenyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, indolyl, benzofuranyl, benzothiophenyl, benzopyrazolyl, benzimidazolyl, benzoxazolyl, benzoisoxazolyl, benzothiazolyl, benzoisothiazolyl, oxopyranyl(pyranonyl), benzopyronyl, coumarinyl, and the like.

The term "heterocycloalkyl" or "heterocyclyl" refers to a saturated or partially unsaturated cyclic hydrocarbon group containing at least one heteroatom. The heterocyclyl ring group can be a monocyclic ring group, a bicyclic ring group, or a tricyclic ring group. The bicyclic ring group can be a spiro-ring group, a bridged-ring group, or a fused-ring group. The heterocyclyl ring group can contain 3 to 20, 3 to 10, 3 to 8, 3 to 7, 5 to 7, 4 to 6, or 5 to 6 ring atoms. The heteroatoms can be one or more selected from the group consisting of N, O and S, for example, 1, 2 or 3 heteroatoms. For example, the heterocyclyl can be pyranyl, aziridinyl, oxiranyl, oxetanyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, piperidinyl, tetrahydropyranyl, dihydropyranyl, morpholinyl, thiomorpholinyl, piperazinyl, or oxazolidinyl.

The term “fused benzoheterocyclyl” refers to a substituent wherein a benzene ring is fused to a heterocyclyl. The fused benzoheterocyclyl may be a fused heterobicyclic ring group in which two adjacent carbon atoms in the ring of said heterocyclyl are shared with the benzene ring. The fused benzoheterocyclyl may be a 9- to 11-membered fused heterobicyclic ring group in which a benzene ring is fused to a 5- to 7-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S. For example, the fused benzoheterocyclyl may include benzopyranyl in which a benzene ring is fused to pyranyl.

The term "fused heterocycloaryl" refers to a substituent having a heterocyclic ring fused to an aryl. A fused heterocycloaryl can be a fused heterobicyclic ring group wherein two adjacent carbon atoms of said heterocyclic ring are shared with said aryl. A fused heterocycloaryl can be a 9- to 7-membered fused heterobicyclic ring group wherein a 5- to 7-membered heterocyclic ring comprising 1 to 3 heteroatoms selected from O, N or S is fused to an aryl. For example, a fused heterocycloaryl can include benzodioxolyl, wherein a dioxolane ring is fused to phenyl.

The above heteroatom may be at least one selected from the group consisting of N, O, P, and S. The above heteroatom may be one, two, or three heteroatoms selected from the group consisting of N, O, and S.

The term "substitution" in "substituted or unsubstituted" refers to the introduction of a substituted hydrogen atom in the case where one or more hydrogen atoms in an organic compound are replaced with another atomic group to form a derivative, and "substituent" refers to the introduced atomic group. As used herein, the term "substituted" without limitation of a substituent includes, for example, a halogen atom, a C ₁ -C ₂₀ alkyl substituted with a halogen atom (e.g., CCF ₃ , CHCF ₂ , CH ₂ F, CCl ₃ , etc.), a C ₁ -C ₂₀ alkoxy, a C ₂ -C ₂₀ alkoxyalkyl, a hydroxy group, -NH ₂ , =NH, nitro, cyano, amidino, hydrazine, hydrazone, carboxy or a salt thereof, sulfonyl, sulfamoyl, a sulfonic acid group or a salt thereof, phosphoric acid or a salt thereof, C ₁ -C ₂₀ alkyl, C ₂ -C ₂₀ alkenyl, C ₂ -C ₂₀ alkynyl, C ₆ -C ₂₀ aryl, C ₆ -C ₂₀ arylalkyl, C ₆ -C ₂₀ heteroaryl, C ₇ -C ₂₀ It may mean substituted with heteroarylalkyl, C ₆ -C ₂₀ heteroaryloxy, C ₆ -C ₂₀ heteroaryloxyalkyl, or C ₆ -C ₂₀ heteroarylalkyl.

The "isomer" of the term "stereoisomer" refers to a compound that has the same molecular formula but has different ways of connecting or arranging the atoms in the molecule. Isomers include, for example, structural isomers and stereoisomers. The stereoisomers can be diastereomers or enantiomers. Enantiomers are isomers that do not overlap with their mirror images, like the relationship between left and right hands, and are also called optical isomers. Enantiomers are distinguished as R (Rectus: clockwise) and S (Sinister: counterclockwise) when four or more substituents are different at the chiral center carbon. Diastereomers are stereoisomers that are not mirror images, and are isomers that occur due to different spatial arrangements of atoms. The above diastereomers can be divided into cis-trans isomers and conformational isomers (or conformers).

The term "solvate" refers to a compound that is solvated in an organic or inorganic solvent. The solvate is, for example, a hydrate.

The term "salt" refers to inorganic and organic acid addition salts of a compound. The pharmaceutically acceptable salt may be a salt which does not cause significant irritation to the organism to which the compound is administered and does not impair the biological activity and physical properties of the compound. The inorganic acid salt may be a hydrochloride, a bromate, a phosphate, a sulfate, or a disulfate. The organic acid salt may be a formate, an acetate, a propionate, a lactate, an oxalate, a tartrate, a malate, a maleate, a citrate, a fumarate, a besylate, a camsylate, an edisyl salt, a trichloroacetic acid, a trifluoroacetate, a benzoate, a gluconate, a methanesulfonate, a glycolate, a succinate, a 4-toluenesulfonate, a galacturonate, an emboxide, a glutamate, an ethanesulfonate, a benzenesulfonate, a p-toluenesulfonate, or an aspartate. The above metal salt may be a calcium salt, a sodium salt, a magnesium salt, a strontium salt, or a potassium salt.

The compound of the above chemical formula 1 may be an agent for SOX9 (SRY-Box Transcription Factor 9). SOX9 recognizes the CCTTGAG sequence together with other members of the HMG-box family of DNA binding proteins, and is known to be expressed by proliferation of non-hypertrophic chondrocytes essential for differentiation of precursor cells into chondrocytes. The compound of the above chemical formula 1 may induce aggregation of the SOX9 transcription factor, thereby increasing the expression of SOX9. The compound of the above chemical formula 1 may be an aggregation inducer of the SOX9 transcription factor, a SOX9 activator, or an activator.

One aspect of the present invention provides a pharmaceutical composition comprising the compound, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof.

One aspect of the present invention provides a pharmaceutical composition for preventing or treating a disease associated with decreased expression of SOX9 (SRY-Box Transcription Factor 9), comprising the compound, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof.

The compounds, stereoisomers, solvates, pharmaceutically acceptable salts, and SOX9 are as described above.

The diseases associated with the above-mentioned decrease in SOX9 expression may be due to the inhibition of SOX9.

In some embodiments, the diseases associated with decreased expression of SOX9 may be osteoarthritis and osteoarthritis-related diseases.

In some embodiments, the osteoarthritis associated condition may be chondropathia, osteonecrosis, or chronic pain.

The term "prevention" refers to any action that inhibits the occurrence of a disease associated with SOX9 or delays its onset by administering the pharmaceutical composition. The term "treatment" refers to any action that improves or beneficially changes the symptoms of a disease associated with SOX9 by administering the pharmaceutical composition.

The pharmaceutical composition may include a pharmaceutically acceptable carrier. The carrier is used to mean an excipient, a diluent or an auxiliary. The carrier may be selected from the group consisting of, for example, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, polyvinyl pyrrolidone, water, saline, a buffer such as PBS, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate, and mineral oil. The composition may include fillers, anticoagulants, lubricants, wetting agents, flavoring agents, emulsifiers, preservatives, or a combination thereof.

The pharmaceutical composition described above can be prepared in any dosage form according to a conventional method. The composition can be formulated, for example, as an oral dosage form (e.g., a powder, a tablet, a capsule, a syrup, a pill, or a granule), or a parenteral dosage form (e.g., an injection). In addition, the composition can be prepared as a systemic dosage form, or a topical dosage form.

In the pharmaceutical composition, the solid preparation for oral administration may be a tablet, a pill, a powder, a granule, or a capsule. The solid preparation may further contain an excipient. The excipient may be, for example, starch, calcium carbonate, sucrose, lactose, or gelatin. In addition, the solid preparation may further contain a lubricant, such as magnesium stearate or talc. In the pharmaceutical composition, the liquid preparation for oral administration may be a suspension, an oral solution, an emulsion, or a syrup. The liquid preparation may contain water or liquid paraffin. The liquid preparation may contain an excipient, for example, a wetting agent, a sweetener, a flavoring agent, or a preservative. In the pharmaceutical composition above, the preparation for parenteral administration may be a sterile aqueous solution, a non-aqueous solution, a suspension, an emulsion, a lyophilized product or a suppository. The non-aqueous solution or the suspension may contain a vegetable oil or an ester. The vegetable oil may be, for example, propylene glycol, polyethylene glycol, or olive oil. The ester may be, for example, ethyl oleate. The base of the suppository may be witepsol, macrogol, Tween 61, cacao butter, laurin butter, or glycerogelatin.

The pharmaceutical composition comprises a compound according to one aspect, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient of the pharmaceutical composition. The term "active ingredient" refers to a physiologically active substance used to achieve pharmacological activity (e.g., treatment of a disease associated with decreased expression of SOX9).

The pharmaceutical composition may contain an effective amount of a compound according to one aspect, a stereoisomer thereof, a solvate, or a pharmaceutically acceptable salt thereof. The term "effective amount" refers to an amount sufficient to exhibit an effect of preventing or treating a disease when administered to a subject in need of prevention or treatment. The effective amount may be appropriately selected by a person skilled in the art depending on the cell or subject selected. A preferred dosage of the pharmaceutical composition may vary depending on the condition and weight of the subject, the extent of the disease, the drug form, the route and duration of administration, but may be appropriately selected by a person skilled in the art. The effective amount may be about 0.5 μg to about 2 g, about 1 μg to about 1 g, about 10 μg to about 500 mg, about 100 μg to about 100 mg, or about 1 mg to about 50 mg per pharmaceutical composition. However, the compound, its stereoisomer, solvate, or pharmaceutically acceptable salt may be administered in divided doses of, for example, about 0.0001 mg/kg to about 100 mg/kg, or about 0.001 mg/kg to about 100 mg/kg, once to 24 times a day, once to 7 times every 2 days to 1 week, or once to 24 times every 1 month to 12 months. In the pharmaceutical composition, the compound, its stereoisomer, solvate, or pharmaceutically acceptable salt may be included in an amount of about 0.0001 wt % to about 10 wt %, or about 0.001 wt % to about 1 wt %, based on the total weight of the composition.

The route of administration may be oral or parenteral. The route of administration may be, for example, oral, transdermal, subcutaneous, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, topical, intranasal, intratracheal, or intradermal. The composition may be administered systemically or locally, and may be administered alone or in combination with other pharmaceutically active compounds.

One aspect of the present invention provides a method for preventing or treating a disease associated with decreased expression of SOX9, comprising the step of administering to a subject the compound, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof.

The compounds, stereoisomers, solvates, pharmaceutically acceptable salts, SOX9, diseases associated with decreased expression of SOX9, prevention, and treatment are as described above.

The subject may be a mammal, such as a human, a mouse, a rat, a cow, a horse, a pig, a dog, a monkey, a sheep, a goat, an ape, or a cat. The subject may be an individual suffering from, or likely to suffer from, a symptom associated with a disease associated with decreased expression of SOX9.

The method may further comprise a step of administering to the subject a known effective ingredient having an effect of preventing or treating a disease associated with SOX9. The known effective ingredient may be administered to the subject simultaneously, separately, or sequentially with the compound according to one aspect, a stereoisomer, a solvate, or a pharmaceutically acceptable salt thereof.

The route of administration may be oral or parenteral. The route of administration may be, for example, oral, transdermal, subcutaneous, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, topical, intranasal, intratracheal, or intradermal. The pharmaceutical composition may be administered systemically or locally, and may be administered alone or in combination with other pharmaceutically active compounds.

The preferred dosage of the pharmaceutical composition above varies depending on the patient's condition and weight, the extent of the disease, the drug form, the route and period of administration, and can be appropriately selected by those skilled in the art. The dosage may be, for example, in the range of about 0.001 mg/kg to about 100 mg/kg, about 0.01 mg/kg to about 10 mg/kg, or about 0.1 mg/kg to about 1 mg/kg for adults. The administration may be administered once a day, twice to 24 times a day, once to twice every three days, once to six times a week, once to 10 times every two weeks, once to 15 times every three weeks, once to three times every four weeks, or once to 12 times a year.

One aspect of the present invention provides a compound, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof for use in preventing or treating a disease associated with decreased expression of SOX9.

The compounds, stereoisomers, solvates, pharmaceutically acceptable salts, SOX9, diseases associated with decreased expression of SOX9, prevention, and treatment are as described above.

One aspect of the present invention provides the use of a compound, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof, for use in the prevention or treatment of a disease associated with decreased expression of SOX9.

The compounds, stereoisomers, solvates, pharmaceutically acceptable salts, SOX9, diseases associated with decreased expression of SOX9, prevention, and treatment are as described above.

One aspect of the present invention provides a use of a compound according to one aspect, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof, for use in the manufacture of a medicament for preventing or treating a disease associated with decreased expression of SOX9.

The compounds, stereoisomers, solvates, pharmaceutically acceptable salts, SOX9, diseases associated with decreased expression of SOX9, prevention, and treatment are as described above.

According to the compound of the present invention and the pharmaceutical composition containing the compound for preventing or treating a disease associated with decreased expression of SOX9, a disease associated with decreased expression of SOX9 can be effectively prevented or treated.

Figure 1 shows data evaluating the SOX9 transcriptional effect of an example compound through a luciferase assay.

Figure 2a is a histogram showing the distribution of SOX9 transcriptional activation effects of example compounds.

Figure 2b is a box plot graph showing the transcriptional activation effect of SOX9 depending on the aggregation of the example compound.

Figure 3a is a fluorescence image of SOX9 overexpressing cell lysates treated with compound B8 and compound C9 and measured.

Figure 3b is a graph measuring the transcriptional activation effect of SOX9 using the luciferase technique.

Figure 4a is a fluorescence image confirming the degree of SOX9 aggregate formation according to the structure of the drug.

Figure 4b is a graph showing the change in the transcriptional activation effect of SOX9 according to the structure of the drug, measured using the luciferase technique.

Figure 5a is a graph showing the degree of atypicality according to the structure and location of SOX9.

Figure 5b is a graph showing the degree of SOX9 aggregation induced by drugs according to SOX9 deletion mutants.

Figure 5c is a graph showing the degree of drug-induced SOX9 aggregation according to deletion variants of the atypical region of the C-terminus of SOX9.

Figure 6 is a graph showing the degree of SOX9 aggregation induced by drugs according to substitution of aromatic amino acids at the C-terminus of SOX9.

Figure 7a is a schematic flow diagram illustrating a method for identifying proteins in drug-induced aggregates using SOX9-turboID.

Figure 7b is a volcano plot showing proteins identified in large amounts according to drug treatment versus the solvent that dissolves the drug.

Figure 8a is a histogram showing the degree of amorphousness of frequently observed proteins depending on the solvent dissolving the drug and the drug treatment.

Figure 8b is a histogram showing the pattern similarity of aromatic amino acids at the C-terminus of SOX9 with the drug-dissolving solvent and the protein frequently observed with drug treatment.

Figure 8c is a graph showing the relationship between the degree of pattern similarity between the aromatic amino acids at the C-terminus of SOX9 and proteins frequently observed according to the solvent dissolving the drug and drug treatment and the conditional probability.

Figure 9 is an image of SOX9 observed within the nucleus of a cell using a general fluorescence microscope and a super-resolution microscope (dSTORM).

Figure 10 shows the qPCR results measuring the mRNA levels of SOX9 target genes according to drug treatment.

Figure 11 is a fluorescence image of mRNA of SOX9 target genes (Acan, Col9a1, Sox9) measured by FISH together with SOX9 protein.

Figure 12a is a histogram showing the size distribution of SOX9 aggregates near the mRNAs of SOX9 target genes according to drug treatment.

Figure 12b is a graph showing the change in the size of SOX9 aggregates near the mRNA of SOX9 target genes depending on drug treatment.

Figure 13 is a graph showing the extent to which SOX9 binds to regulatory regions of genes in DNA depending on the solvent dissolving the drug and the drug treatment.

Figure 14 is a graph showing the extent to which SOX9 binds to individual gene regulatory regions of DNA depending on the solvent dissolving the drug and the drug treatment.

Figure 15a is a schematic flow chart for an experiment administering drugs to osteoarthritis model mice.

Figure 15b is an image showing the extent to which cartilage tissue in the joint area of an osteoarthritis model mouse was restored according to drug treatment (red: cartilage, blue: bone).

Figure 15c is a graph showing the extent of tissue recovery evaluated using the OARSI grade according to drug treatment.

Figure 16 is a schematic flow chart and graph of the results of an experiment that compared the degree of weight-bearing on the leg of an osteoarthritis model rat treated with drugs with a leg that did not undergo surgery.

Figure 17 is a schematic flow chart and graph of the results of an experiment that quantitatively measured and evaluated the degree to which the sole of an osteoarthritis model rat could withstand a needle according to drug treatment.

The present invention will be described in more detail through the following examples. However, these examples are provided for illustrative purposes only and the scope of the present invention is not limited to these examples.

Manufacturing Example A1: Synthesis of 3-(2-aminothiazol-4-yl)-7-methoxycoumarin

Step 1: 3-Acetyl-7-methoxycoumarin

2-Hydroxy-4-methoxybenzaldehyde (2 mmol, CAS: 673-22-3), ethyl acetoacetate (2.4 mmol), and piperidine (0.2 mmol) were added to ethanol (4 mL) and stirred. After the reaction was completed, the temperature of the reaction product was lowered using an ice water bath, and the precipitate was filtered to obtain 3-acetyl-7-methoxycoumarin in a yield of 88%.

^1H NMR (400 MHz, DMSO) δ 8.64 (s, 1H), 7.87 (d, J = 8.7 Hz, 1H), 7.06 (d, J = 2.3 Hz, 1H), 7.02 (dd, J = 8.7, 2.4 Hz, 1H), 3.90 (s, 3H), 2.56 (s, 3H).

Step 2: 3-Bromoacetyl-7-methoxycoumarin

The obtained product (1 mmol) of Step 1 above and tetrabutylammonium tribromide (TBATB, 2 mmol) were dissolved in CH ₂ Cl ₂ (10 mL) and stirred at room temperature. After the reaction was completed, the resulting precipitate was separated by filtration, thereby obtaining 3-bromoacetyl-7-methoxycoumarin in a yield of 59%.

^1H NMR (400 MHz, DMSO) δ 8.82 (s, 1H), 7.93 (d, J = 8.7 Hz, 1H), 7.12 (d, J = 2.4 Hz, 1H), 7.06 (dd, J = 8.7, 2.5 Hz, 1H), 4.87 (s, 2H), 3.92 (s, 3H).

Step 3: 3-(2-Aminothiazol-4-yl)-7-methoxycoumarin

The obtained product (1 mmol) of step 2 above and thiourea (2 mmol) were added to ethanol (10 mL) and heated with stirring. After the reaction was completed, the resulting precipitate was filtered to obtain intermediate a-1 in 99% yield.

^1H NMR (400 MHz, DMSO) δ 8.47 (s, 1H), 7.75 (d, J = 8.7 Hz, 1H), 7.42 (s, 1H), 7.07 (d, J = 2.4 Hz, 1H), 7.00 (d, J = 8.6 Hz, 1H), 3.88 (s, 3H).

Manufacturing Examples A2 to A13

Intermediates a-2 to a-12 were obtained in the same manner as in Manufacturing Example A1, except that the hydroxybenzaldehyde compounds in Table 1 below were used instead of 2-hydroxy-4-methoxybenzaldehyde in Step 1 of Manufacturing Example A1.

[Table 1]

Manufacturing Example A13: Synthesis of 3-(2-aminooxazol-4-yl)-7-methoxycoumarin

Intermediate a-13 (52%, 387 mg) was obtained in the same manner as in Manufacturing Example A1, except that urea (CAS: 57-13-6) was used instead of thiourea in step 3 of Manufacturing Example A1.

^1H NMR (300 MHz, DMSO) δ 8.24 (s, 1H), 7.84 (s, 1H), 7.77 (d, J = 8.7 Hz, 1H), 7.05 (d, J = 2.6 Hz, 1H), 6.98 (dd, J = 8.7, 2.4 Hz, 1H), 6.83 (s, 2H), 3.87 (s, 3H).

The compounds in the table below were used as intermediates a-14 to a-17.

[Table 2]

Manufacturing Example A18: Synthesis of 4-(2-aminothiazol-4-yl)biphenyl

Intermediate a-18 (94%, 476 mg) was obtained in the same manner as in Step 3 of Preparation A1, except that 2-bromo-4'-phenylacetophenone (CAS: 135-73-9) was used instead of the obtained product of Step 2 of Preparation A1.

¹ H NMR (400 MHz, DMSO) δ 8.85 (s, 2H), 7.88 - 7.78 (m, 4H), 7.78 - 7.69 (m, 2H), 7.50 (dd, J = 8.5, 6.9 Hz, 2H), 7.44 - 7.36 (m, 1H), 7.32 (s, 1H).

Manufacturing Example A19: Synthesis of 3-(2-aminothiazol-4-yl)-6-methyl-2H-pyran-2,3(3H)-dione

Step 1: 3-(2-bromoacetyl)-6-methyl-2H-pyran-2,4(3H)-dione

Dehydroacetic acid (5.0 mmol, CAS: 520-45-6) and p-TsOH (5.5 mmol, CAS: 6192-52-5) were stirred in acetonitrile (10 mL), and NBS (5.5 mmol, CAS: 128-08-5) was slowly added and heated. After 9 h, water was added to the reaction mixture, and the product was extracted with DCM and concentrated to obtain the product (see Synthetic Communications, 2012, 42(18), 2739-2747).

Step 2: 3-(2-Aminothiazol-4-yl)-6-methyl-2H-pyran-2,3(3H)-dione

Using the obtained product of step 1 above, intermediate a-19 (8%, 94 mg) was obtained in the same manner as step 3 of Manufacturing Example A1.

^1H NMR (400 MHz, DMSO) δ 15.12 (s, 1H), 8.02 (s, 2H), 7.10 (s, 1H), 6.12 (d, J = 1.0 Hz, 1H), 2.20 (d, J = 0.9 Hz, 3H).

Manufacturing Example A20: Synthesis of (2-aminothiazol-5-yl)(phenyl)methanone

Step 1: (E)-3-(dimethylamino)-1-phenylprop-2-en-1-one

Acetophenone (2.0 mmol) and N,N-dimethylformamide dimethylacetal (4.0 mmol) were stirred and heated in toluene (2 mL) for 18 h, then the solvent was removed and (E)-3-(dimethylamino)-1-phenylprop-2-en-1-one (124 mg, 35%) was isolated by column chromatography (DCM/MeOH=5:1).

Step 2: (2-Aminothiazol-5-yl)(phenyl)methanone

The obtained product (0.4 mmol) of Step 1, S powder (1.7 mmol), cyanamide solution (1.7 mmol), and N-methylporpholine (NMM, 0.1 mmol) were stirred in N-methylpyrrolidone (NMP, 1 mL) and reacted at 100 °C for 16 h under an argon atmosphere. After that, the reaction mixture was cooled, diluted with EA, and extracted by adding water. The organic layer including the product was removed with Na ₂ SO ₄ to remove the remaining moisture, concentrated, and purified through column chromatography (Hex / EA = 1:1) to obtain intermediate a-20 (31%, 78 mg) (see J. Org. Chem. 2019, 84(18), 12237-12245).

^1H NMR (600 MHz, CDCl3) δ 7.78 (dd, J = 8.3, 1.4 Hz, 2H), 7.62 (s, 1H), 7.60 - 7.56 (m, 1H), 7.53 - 7.46 (m, 2H), 6.22 (br s, 2H).

Manufacturing Example B1: Synthesis of 3-(2-succinimidylthiazol-4-yl)-7-methoxycoumarin

Anhydrous succinic acid (10 mmol, 1.00 g) and intermediate a-1 (1 mmol) were placed in a mortar and mixed well, and then transferred to a reactor. The reaction mixture was stirred at 150 ℃ for 5 hours. After the reaction was completed, the obtained product was purified by silica gel chromatography (CH ₂ Cl ₂ : CH ₃ C(O)CH ₃ = 20:1) to obtain intermediate b-1 (yield: 84%, 300 mg).

^1H NMR (400 MHz, DMSO) δ 8.65 (s, 1H), 8.39 (s, 1H), 7.84 (d, J = 8.7 Hz, 1H), 7.09 (d, J = 2.4 Hz, 1H), 7.00 (dd, J = 8.7, 2.4 Hz, 1H), 3.88 (s, 3H), 2.87 (s, 4H).

Manufacturing examples B2 to B21

In Manufacturing Example B1, intermediate a-1 was changed to intermediates a-2 to a-20 to synthesize intermediates b-2 to b-20, respectively.

[Table 3]

Example 1: N ¹ -(3,4-dimethoxyphenethyl)-N ⁴ - Synthesis of (4-(7-methoxycoumarin-3-yl)thiazol-2-yl)succinamide

Intermediate b-1 (0.20 mmol) was dissolved in anhydrous dioxane (2 mL) under argon gas conditions in a reactor equipped with a condenser, and 2-(3,4-dimethoxyphenyl)ethan-1-amine (0.22 mmol (CAS: 120-20-7, TCI/D0678)) was added. The reaction mixture was stirred at 110°C for 5 hours. After the reaction was completed, the reaction mixture was concentrated under vacuum. Water (20 mL) was added to the obtained residue to precipitate the product. The precipitated product was separated by centrifugation or filtration and lyophilized to obtain the compound of Example 1 (Compound B8, yield: 89%, 96 mg).

^1H NMR (400 MHz, DMSO) δ 12.30 (s, 1H), 8.54 (s, 1H), 7.99 (t, J = 5.6 Hz, 1H), 7.89 (s, 1H), 7.77 (d, J = 8.7 Hz, 1H), 7.09 (d, J = 2.4 Hz, 1H), 7.00 (dd, J = 8.6, 2.5 Hz, 1H), 6.84 (d, J = 8.2 Hz, 1H), 6.80 (d, J = 2.0 Hz, 1H), 6.70 (dd, J = 8.1, 2.0 Hz, 1H), 3.88 (s, 3H), 3.74 (s, 3H), 3.70 (s, 3H), 3.23 (dt, J = 6.7, 6.7 Hz, 2H), 2.68 (t, J = 6.9 Hz, 2H), 2.62 (t, J = 7.4 Hz, 2H), 2.44 (t, J = 7.0 Hz, 2H).

Examples 2 to 81

Compounds of Examples 2 to 81 were obtained in the same manner as in Example 1, except that intermediate b-1 and 2-(3,4-dimethoxyphenyl)ethan-1-amine of Example 1 were changed as follows.

Example number Intermediate b Amine compounds Example 2 Intermediate b-1 3,4-Dimethoxybenzylamine (CAS: 5763-61-1, Alfa Aesar/B25033) Example 3 Intermediate b-1 2-(2-Aminoethyl)pyridine (CAS: 2706-56-1, TCI/A1999) Example 4 Intermediate b-1 2-(4-Aminophenyl)ethylamine (CAS: 13472-00-9, Alfa Aesar/L15782) Example 5 Intermediate b-1 L-Phenylalanine methyl ester hydrochloride (CAS: 7524-50-7, TCI/P1278) Example 6 Intermediate b-1 4-(2-Aminoethyl)morpholine (CAS: 2038-03-1, Sigma/A55004) Example 7 Intermediate b-1 2-Phenylethylamine (CAS: 64-04-0, TCI/P0085) Example 8 Intermediate b-1 Dopamine hydrochloride (CAS: 62-31-7, Alfa/A11136) Example 9 Intermediate b-1 3-(2-Aminoethyl)pyridine (CAS: 20173-24-4, Alfa/L16311) Example 10 Intermediate b-1 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 11 Intermediate b-1 Histamine (CAS: 51-45-6, Alfa/J61727) Example 12 Intermediate b-1 2-(4-nitrophenyl)ethylamine hydrochloride (CAS: 29968-78-3, Alfa/H64894) Example 13 Intermediate b-1 2-(2-Aminoethyl)thiophene (CAS: 30433-91-1, TCI/A1637) Example 14 Intermediate b-1 2-(3-methoxyphenyl)ethylamine (CAS: 2039-67-0, TCI/M1891) Example 15 Intermediate b-1 2-(4-methoxyphenyl)ethylamine (CAS: 55-81-2, TCI/M0795) Example 16 Intermediate b-1 3-Phenylpropylamine (CAS: 2038-57-5, TCI/P0664) Example 17 Intermediate b-1 Benzylamine (CAS:100-46-9, TCI/B0406) Example 18 Intermediate b-1 2,2-Diphenylethylamine (CAS: 3963-62-0, TCI/D2018) Example 19 Intermediate b-1 Tryptamine (CAS: 61-54-1, Alfa Aesar/A11116.09) Example 20 Intermediate b-1 2-(3,4-(methylenedioxy)phenyl)ethylamine hydrochloride (CAS: 1653-64-1, Alfa Aesar/H60208) Example 21 Intermediate b-1 2-Naphthaleneethanamine (CAS: 2017-68-7, Sigma/667167) Example 22 Intermediate b-1 2-(3,4-Dichlorophenyl)ethylamine (CAS: 21581-45-3, TCI/D2927) Example 23 Intermediate b-1 2-(p-Tolyl)ethylamine (CAS: 3261-62-9, Alfa Aesar/15788) Example 24 Intermediate b-1 2-(2-Fluorophenyl)ethylamine (CAS: 52721-69-4, TCI/F0933) Example 25 Intermediate b-1 2-(4-Fluorophenyl)ethylamine (CAS: 1583-88-6, TCI/F0829) Example 26 Intermediate b-1 4-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride (CAS: 30827-99-7, Alfa Aesar/H26473) Example 27 Intermediate b-1 N-(2-Aminoethyl)piperazine (CAS: 140-31-6, TCI/A0304) Example 28 Intermediate b-1 4-(2-Aminoethyl)thiomorpholine 1,1-dioxide (CAS: 89937-52-0, TCI/A1803) Example 29 Intermediate b-1 4-Picolylamine (CAS: 3731-53-1, TCI/P0779) Example 30 Intermediate b-1 3-(Pyridin-4-yl)propan-1-amine (CAS:30532-36-6, Ambeed/A139358) Example 31 Intermediate b-1 Pyrimidin-4-ylmethanamine dihydrochloride (CAS: 618446-08-5, Ambeed/A283503) Example 32 Intermediate b-1 Pyridazin-4-ylmethanamine dihydrochloride (CAS: 1028615-75-9, Ambeed/ A524320) Example 33 Intermediate b-1 (2-Fluoropyridin-4-yl)methanamine dihydrochloride (CAS: 667906-60-7, Ambeed/A210034)) Example 34 Intermediate b-1 (2-Chloropyridin-4-yl)methanamine hydrochloride (CAS: 916210-98-5, Ambeed/A166155)) Example 35 Intermediate b-1 (2-Bromopyridin-4-yl)methanamine (CAS: 858362-82-0, Ambeed/ A150324) Example 36 Intermediate b-1 4-(Aminomethyl)pyridin-2-amine (CAS: 199296-51-0, Ambeed/A548509) Example 37 Intermediate b-1 Pyridine-2,4-diamine (CAS: 461-88-1, Ambeed/A202971) Example 38 Intermediate b-1 3,4,5-Trimethoxybenzylamine (CAS: 18638-99-8, TCI/T3087) Example 39 Intermediate b-1 8-Aminoquinoline (CAS: 578-66-5, Alfa/C12371) Example 40 Intermediate b-14 2-(3,4-Dimethoxyphenyl)ethylamine (CAS: 120-20-7, TCI/D0678) Example 41 Intermediate b-15 2-(3,4-Dimethoxyphenyl)ethylamine (CAS: 120-20-7, TCI/D0678) Example 42 Intermediate b-16 2-(3,4-Dimethoxyphenyl)ethylamine (CAS: 120-20-7, TCI/D0678) Example 43 Intermediate B-17 2-(3,4-Dimethoxyphenyl)ethylamine (CAS: 120-20-7, TCI/D0678) Example 44 Intermediate b-4 2-(3,4-Dimethoxyphenyl)ethylamine (CAS: 120-20-7, TCI/D0678) Example 45 Intermediate b-5 2-(3,4-Dimethoxyphenyl)ethylamine (CAS: 120-20-7, TCI/D0678) Example 46 Intermediate b-3 2-(3,4-Dimethoxyphenyl)ethylamine (CAS: 120-20-7, TCI/D0678) Example 47 Intermediate b-7 2-(3,4-Dimethoxyphenyl)ethylamine (CAS: 120-20-7, TCI/D0678) Example 48 Intermediate b-8 2-(3,4-Dimethoxyphenyl)ethylamine (CAS: 120-20-7, TCI/D0678) Example 49 Intermediate b-9 2-(3,4-Dimethoxyphenyl)ethylamine (CAS: 120-20-7, TCI/D0678) Example 50 Intermediate b-6 2-(3,4-Dimethoxyphenyl)ethylamine (CAS: 120-20-7, TCI/D0678) Example 51 Intermediate b-10 2-(3,4-Dimethoxyphenyl)ethylamine (CAS: 120-20-7, TCI/D0678) Example 52 Intermediate b-2 2-(3,4-Dimethoxyphenyl)ethylamine (CAS: 120-20-7, TCI/D0678) Example 53 Intermediate b-20 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 54 Intermediate b-20 2-(3,4-Dimethoxyphenyl)ethylamine (CAS: 120-20-7, TCI/D0678) Example 55 Intermediate b-4 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 56 Intermediate b-5 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 57 Intermediate b-3 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 58 Intermediate b-7 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 59 Intermediate b-6 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 60 Intermediate b-10 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 61 Intermediate b-2 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 62 Intermediate b-9 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 63 Intermediate b-11 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 64 Intermediate b-12 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 65 Intermediate b-14 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 66 Intermediate b-15 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 67 Intermediate b-16 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 68 Intermediate B-17 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 69 Intermediate b-13 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 70 Intermediate b-13 2-(3,4-Dimethoxyphenyl)ethylamine (CAS: 120-20-7, TCI/D0678) Example 71 Intermediate B-18 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 72 Intermediate b-20 4-(2-Aminoethyl)pyridine (CAS: 13258-63-4, TCI/A1264) Example 73 Intermediate b-20 2-(3,4-Dimethoxyphenyl)ethylamine (CAS: 120-20-7, TCI/D0678) Example 74 Intermediate b-1 5,6-Dimethoxy-2-(4-piperidylmethyl)-1-indanone hydrochloride (CAS: 120013-39-0, TCI/D3997) Example 75 Intermediate b-1 4-(4-Aminopiperidino)pyridine dihydrochloride (CAS: 1169396-92-2, Ambeed/A689438) Example 76 Intermediate b-1 1-(4-Pyridyl)piperazine (CAS: 1008-91-9, TCI, P1863) Example 77 Intermediate b-1 4-(4-piperidyl)pyridine (CAS: 581-45-3, TCI/P2012) Example 78 Intermediate b-1 Trimetazidine dihydrochloride (CAS: 13171-25-0, TCI/T2726) Example 79 Intermediate b-1 Quinoline-4-carbohydrazide (CSA: 29620-62-0, Ambeed/A600347) Example 80 Intermediate b-1 1H-Indole-6-carbohydrazide (CAS: 851211-74-0, Ambeed/A208763) Example 81 Intermediate b-1 5-(Pyridin-4-yl)-1,3,4-thiadiazol-2-amine (CAS: 2002-04-2, Ambeed/A857207)

[Table 5]

Experimental Example 1: Evaluation of SOX9 transcriptional effect activity

To determine whether the compound enhances SOX9 transcriptional effector activity, a luciferase assay capable of measuring the transcriptional activity of SOX9 was performed using the following experimental method.

Primary rat chondrocytes were extracted from rat cartilage. The extracted cells were stabilized in a low-oxygen environment ( _O2 3%, CO2 ₅ %) incubator. At this time, the medium used was DMEM (Dulbecco Modified Eagle Medium, LM001-05, Welgene) supplemented with 10% FBS (fetal bovine serum, Gibco).

Cells were stabilized and treated with hyaluronidase type I-S (Sigma-Aldrich, Hase) for 4 h for transfection of the luciferase plasmid.

For transfection, Metafectene pro reagent (Biontex) was used to treat cells with the 4X48pCol2a1-Luicferase plasmid, whose expression is promoted by the transcriptional activity of SOX9, and the Renilla luciferase plasmid, which continuously expresses the plasmid for the control for transfection, for 6 h. The 4X48pCol2a1-Luicferase plasmid and the Renilla luciferase plasmid were used as described in Kim, S. et al. “Tankyrase inhibition preserves osteoarthritic cartilage by coordinating cartilage matrix anabolism via effects on SOX9 PARylation”. Nat Commun 10, 4898 (2019).

After 6 hours, the medium was replaced with a culture medium containing the compounds. At this time, each compound was dissolved in DMSO at a concentration of about 5 mM to 10 mM and used in the medium at a concentration of 7.5 mM.

After 48 hours, the cells were lysed using a Dual Luciferase Assay Kit (Promega) and reacted with a luciferase substrate, and the luminescence from luciferase (relative to DMSO) was measured using a microplate reader, and the results are shown in Table 6 and the graph in Fig. 1 according to the following criteria.

++++: 8 or more / +++: less than 8, 5 or more / ++: less than 5, 2 or more / +: less than 2, more than 1

Example Compound number SOX9 activation Example Compound number SOX9 activation Example Compound number SOX9 activation 1 B8 ++ 27 C26 +++ 53 D14 ++ 2 C1 ++++ 28 C27 + 54 D15 ++ 3 C2 ++ 29 C28 ++++ 55 D16 ++ 4 C3 ++ 30 C29 +++ 56 D17 ++ 5 C4 ++ 31 C30 ++ 57 D18 ++ 6 C5 +++ 32 C31 ++ 58 D19 ++ 7 C6 ++++ 33 C32 ++ 59 D20 +++ 8 C7 ++++ 34 C33 ++ 60 D21 ++ 9 C8 ++++ 35 C34 ++ 61 D22 ++ 10 C9 ++++ 36 C35 ++ 62 D23 ++ 11 C10 ++ 37 C36 ++ 63 D24 ++ 12 C11 ++ 38 C37 +++ 64 D25 ++ 13 C12 ++ 39 C38 ++++ 65 D26 + 14 C13 ++ 40 D1 + 68 D29 +++ 15 C14 ++ 41 D2 + 69 D30 + 16 C15 ++ 42 D3 + 70 D31 ++ 17 C16 ++ 43 D4 ++ 71 D32 + 18 C17 ++ 44 D5 ++ 74 C43 ++ 19 C18 ++ 45 D6 ++ 75 C39 +++ 20 C19 ++ 46 D7 ++ 76 C40 +++ 21 C20 ++ 47 D8 ++ 77 C41 ++ 22 C21 ++ 48 D9 ++ 79 C44 ++++ 23 C22 ++ 49 D10 ++ 80 C45 ++++ 24 C23 + 50 D11 ++ 81 C46 ++ 25 C24 ++++ 51 D12 ++ 26 C25 ++ 52 D13 ++

Referring to Table 6 and Figure 1, it was confirmed that the compound of the present invention has an excellent SOX9 transcriptional activation effect.

The compounds of the examples showed different effects in increasing the degree of transcriptional activity (Fig. 2a), and when checking the change in the degree of transcriptional activity depending on whether or not the aggregation phenomenon was induced, it was found that there was a statistically significant correlation between the aggregation phenomenon and the transcriptional activity effect (Fig. 2b). This means that the aggregation induction of the drug at the SOX9 protein level affects and is related to the drug efficacy.

Experimental Example 2: Induction of SOX9 aggregation phenomenon and evaluation of transcriptional activity

^9x105 cells to be transfected were plated on a 100 mm dish and grown in a cell incubator for two days. The grown cells were transfected with 7 ug of the SOX9-eGFP plasmid that can be overexpressed in cells using a transfection agent (Poly-jet). At this time, the cells were exposed to DMEM medium without FBS for 30 minutes, and after 30 minutes, the SOX9-eGFP plasmid and Polyjet were added to DMEM, mixed together, and the medium that had been waited for 10 minutes was added. After reacting in the incubator for 4 hours, DMEM medium containing FBS was added and kept in the incubator for two days. Afterwards, the cells were removed using a scraper, centrifuged, and only the cell pellet was frozen in liquid nitrogen and stored in a -80℃ freezer.

The cell lysis buffer containing TritonX100 0.2%, EDTA 1 mM, Hepes 50 mM, NaCl 150 mM, glycerol 10%, protease inhibitor cocktail 1%, phosphatase inhibitor cocktail (tyrosine, serine/threonine) 1%, and TCEP 2 mM was added to the cell pellet to lyse the pellet. The lysed cells were lysed using an ultrasonic homogenizer, and the lysate was centrifuged (15,000 g, 10 min) to obtain the supernatant. The total protein concentration of the obtained supernatant was measured using a DC (detergent-compatible) assay, and the SOX9-eGFP concentration was measured using a fluorometer. The cell lysates whose concentrations were measured were used immediately or frozen in liquid nitrogen and stored at -80°C.

SOX9, which is relatively difficult to purify from E. coli, was expressed and purified from HEK293T using the Halo-Tag purification method. Even when SOX9, not a soluble form, was used, the correlation between the induction of aggregation and the degree of transcriptional activity was repeatedly observed, confirming the validity of using a soluble form (Fig. 2b).

The compound of the example was diluted to 1/100 of the concentration to be reacted and then transferred to a tube according to each reaction volume. The prepared cell lysate was added to the tube to which compound B8 or compound C9 was transferred and reacted. The lysate reacted with the drug was transferred to a 96-well plate with a cover glass surface and waited for 1 hour at room temperature until the reaction was completed. The 96-well plate where the reaction was completed was transferred to a confocal fluorescence microscope and fluorescence imaging was performed.

The captured fluorescence images were analyzed using a self-developed MATLAB code. K-mean clustering was used to distinguish between aggregates and non-aggregates, and the fluorescence intensity, area, shape, and number of aggregates were analyzed.

As a result, it was verified that compounds B8 and C9 induced the aggregation of SOX9 compared to DMSO (Fig. 3a). In addition, using a Luciferase plasmid that specifically reacts to the transcriptional activity of SOX9, it was confirmed that compounds B8 and C9, which induce the aggregation, enhanced the activity of the SOX9 transcription factor (Fig. 3b). In particular, it was found that compound B8 significantly enhanced the transcriptional activity by about 8 times, while compound C9 significantly enhanced the transcriptional activity by about 2 times. At this time, the SOX9 aggregation phenomenon disappeared due to the structural change of compound C9, such as a change in the position or substitution of a nitrogen atom, and it was confirmed that the induction of such aggregation affected the regulation of transcriptional activity (Figs. 4a, 4b). This means that the induction of aggregation and the regulation of transcriptional activity occur based on the chemical structure of the compound.

Experimental Example 3: Evaluation of the degree of aggregation according to the deletion site of SOX9

We investigated the mechanism by which compound C9 contributes to enhancing the osteoarthritis effect.

Specifically, the binding site of the drug was explored based on compound C9, which was confirmed to have the best efficacy. In the case of SOX9, it is distinguished into a structured site and a structureless site, and in particular, the structureless site, IDR, is large enough to account for about 80% of the protein (Fig. 5a). In order to find out which site the aggregation-inducing phenomenon of compound C9 occurs through interaction with, SOX9 mutants with each site deleted were produced. In the same manner as in the above experimental examples, cell lysates for SOX9 mutants were produced, and it was confirmed that the aggregation phenomenon was absent when C9 was deleted in the IDR region corresponding to the C-terminus of the protein (Fig. 5b). Furthermore, in order to narrow down the binding site, mutants with 35 amino acid intervals deleted from the front to the back of the C-terminal region were produced, and cell lysates were produced and confirmed. At this time, it was confirmed that the aggregation phenomenon was drastically reduced when the site corresponding to regions 8 and 9 where aromatic amino acids are gathered was deleted (Fig. 5c).

Experimental Example 4: Changes in the degree of SOX9 aggregation according to mutations in aromatic amino acids at positions 8 and 9 of the C-terminus of SOX9

The importance of aromatic amino acids was confirmed by producing cell lysates expressing mutants in which aromatic amino acids were substituted with alanine. Similar to the mutants lacking regions 8 and 9, mutations in aromatic amino acids corresponding to regions 8 and 9 reduced the aggregation-induced phenomenon by compound C9 (Fig. 6). Through this, it was confirmed that compound C9 induces the aggregation of SOX9 through interaction with the region where aromatic amino acids are gathered in the structureless region of the SOX9 C-terminus.

Experimental Example 5: Identification of proteins within aggregate-induced SOX9 aggregates

In the case of aggregates, they are often created by multiple proteins rather than a single protein. Therefore, in order to identify the proteins that aggregate with SOX9 by treatment with compound C9, SOX9 was labeled with TurboID, and biotin was attached to proteins within 10 nm of SOX9 by treatment with compound C9. The biotin-bound proteins were separated using Streptavidin-Beads, and liquid chromatography-mass spectrometry (LC/MS) was performed (Fig. 7a). Each mass spectrometry experiment was repeated three times for one condition. Through mass spectrometry, proteins such as Pol2, as YTHDF3, CCAR2, MBNL1, NDUFA10, MYO1C, TAF5, and chromatin remodelers (KAT6A, KAT8), which are statistically significantly more frequently observed in the group treated with C9 compared to DMSO, were detected, and proteins known to act together with SOX9, such as SOX5 and SOX6, were also detected (Fig. 7b).

Experimental Example 6: Confirmation of protein atypicality and evaluation of similarity with SOX9-C-IDR through spatial frequency analysis

The sequence information of proteins significantly detected in C9 and DMSO was used to identify the IDR region using Metapredict. At this time, it was confirmed that the proteins detected compared to DMSO had many IDRs according to compound C9 treatment (Fig. 8a). Since IDR is known to be a region that plays an important role in aggregation, it indirectly suggests that compound C9 induces SOX9 aggregation.

Since compound C9 induced aggregation by targeting aromatic amino acids of SOX9 IDR, it was confirmed whether the IDRs of the proteins detected through mass spectrometry contained the site targeted by compound C9. A one-dimensional binary vector was created in which the aromatic amino acids present in the IDRs of the proteins were 1 and the other amino acids were 0, and this was wavelet transformed. The transformed information includes information on the degree of clustering of aromatic amino acids according to time (position) and frequency. The degree of similarity to the C-IDR of SOX9 was compared through cross-correlation analysis between the wavelet transformed data of the proteins detected in this way and the data corresponding to the C-IDR of SOX9. Through this, it was found that the IDRs with a similar pattern to the SOX9 C-IDR in the sample treated with compound C9 had a high probability of having a high similarity score (Fig. 8b). In order to exclude the possibility that the number of proteins with high similarity to the SOX9 C-IDR pattern was originally small, the conditional probability was calculated by considering the number of proteins corresponding to each similarity score. Through this, it was found that IDRs with high similarity scores were more likely to be found in samples treated with compound C9 (Fig. 8c). These results support the fact that compound C9 specifically induces aggregate formation in a specific pattern (spatial frequency) created by aromatic amino acids in the C-IDR of SOX9.

Experimental Example 7: Analysis of SOX9 aggregates using super-resolution fluorescence microscopy dSTORM

To investigate the effect of drugs on the changes in SOX9 aggregates within cells, SOX9 was labeled using IF (Immunofluorescence staining) and fluorescence imaging was performed. Since transcription factor aggregates such as SOX9 aggregates have a size of 100 nm to 1 μm, changes in SOX9 aggregates cannot be confirmed at the level of a general fluorescence microscope. Therefore, dSTORM (direct stochastic optical reconstruction microscopy), one of the super-resolution microscopes, was used to obtain the fluorescence image in Fig. 9.

Experimental Example 8: Evaluation of mRNA levels of genes involved in osteoarthritis and RNA-FISH analysis

To determine the relationship between the observed changes in SOX9 aggregates and transcriptional activity, RNA FISH (Fluorescence In Situ Hybridization) was performed with dSTORM on Col9a1, Acan, and SOX9, which were transcripts whose transcriptional activity was increased by compound C9 and whose mRNA levels were highly increased compared to DMSO, to obtain the mRNA level graph in Fig. 10 and the dSTORM fluorescence image in Fig. 11.

RNA-FISH confirmed that the locations of each RNA foci and SOX9 aggregates observed in the nucleus were well colocalized (Fig. 11). When the size changes of SOX9 aggregates observed near RNA foci were observed in C9 compared to DMSO through image analysis, the size of SOX9 aggregates increased by about 30% to 50% (Fig. 12a, Fig. 12b). This was a high level compared to the 2% change in the size of SOX9 aggregates present in the nucleus. Therefore, it was found that compound C9 is specifically involved in the change in the size of SOX9 aggregates near RNA, which is related to transcriptional activity.

Experimental Example 9: Confirmation of the effect of the compound on the degree of binding between SOX9 and DNA through Cut&Tag

To determine whether the aggregates of SOX9 altered by drugs are actually involved in transcriptional activity, Cut&Tag experiments were performed to identify the DNA site to which SOX9 binds at the genome-wide level. As shown below, the degree to which SOX9 binds to DNA increases with drug treatment (Fig. 13).

Experimental Example 10: Confirmation of changes in the degree of SOX9 binding to the DNA of genes that play a role in bone joint regeneration according to compound treatment

When we checked the information on the degree of DNA binding of SOX9 obtained through Cut&Tag for each gene involved in cartilage production, SOX9, Col2a1, Col9a1, Chad, Acan, and Comp, we could confirm that when treated with C9 drug, SOX9 binding increased throughout the DNA (SOX9, Col2a1, Chad) or increased binding in a specific part of DNA (red box) (Col9a1, Acan, Comp) (data with DMSO subtracted from C9, yellow) compared to when treated with DMSO (Figs. 14a to 14f). These results show that compound C9 affected the size change of SOX9 aggregates and the degree of DNA binding.

Experimental Example 11: Evaluation of the degree of cartilage tissue recovery in a rat model of osteoarthritis

To determine whether compound C9 helps in cartilage production, an experiment was conducted in which compound C9 was injected intra-articularly at a concentration of 750 μM at one-week intervals into a rat model of osteoarthritis (OA) induced by DMM (destabilization of the medial meniscus) surgery for a total of seven times (Fig. 15a). In the case of rats that underwent DMM surgery, the tissues showed damaged cartilage tissue (yellow triangles) rather than recovered when treated with DMSO, whereas the damaged cartilage tissue was recovered when the drug was treated (Fig. 15b). In order to objectively evaluate the recovery of cartilage tissue, when the OARSI Grade (a higher score indicates greater damage to cartilage tissue) was given in a blind test by individual researchers, it was confirmed that the OARSI grade was statistically significantly lower when compound C9 was treated (Fig. 15c). This suggests that compound C9 can help in the recovery of damaged cartilage tissue.

Experimental Example 12: Evaluation of the degree of alleviation of osteoarthritis through behavioral experiments

To confirm the recovery of osteoarthritis at the behavioral level, weight bearing and Von-Frey behavioral experiments were conducted. In the case of the weight bearing experiment, the degree of force loading on the operated or non-operated leg was measured according to the drug treatment, and the experiment was conducted to confirm what changes in the degree of force loading occurred when compound C9 was treated (Fig. 16). When compound C9 was treated, it was confirmed that the level of OA-induced (DMM surgery-treated) mice was almost similar to that of non-induced OA mice, and the degree of change was confirmed to be significant compared to the case of DMSO treatment (Fig. 16).

In the Von-Frey behavioral experiment, a weak stimulus is given to the sole of the rat, and as the stimulus becomes stronger, the intensity of the stimulus at the moment the sole is lifted is measured to evaluate the degree of cartilage recovery of the rat (Fig. 17). As the degree of cartilage recovery improves, the intensity of the stimulus that can be tolerated increases. Compared to rats that are not induced with OA, it was confirmed that the intensity of the stimulus that can be tolerated increases in rats with induced OA according to compound C9 treatment (Fig. 17).

Through these behavioral experimental results, it was confirmed that compound C9 alleviates osteoarthritis at the behavioral level. Therefore, it was verified that the present invention can screen osteoarthritis-causing substances such as compound C9, which can exhibit osteoarthritis-causing effects by aggregating SOX9.

Claims (22)

A compound represented by the following chemical formula 1, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] In the above chemical formula 1, One of Y ¹ and Y ² is N, and the other is S, O, or NR ^a1 ; X ¹ and X ² are each independently O, S or NR ^a2 ; U is NR ⁿ² , a 5- to 7-membered heterocyclyl comprising 1 to 3 heteroatoms N or these linked to each other; Z ¹ is C _1-6 alkylene; Z ² is a direct bond, C _1-6 alkylene, -NR ⁿ³ CO- or a 5- to 7-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O and S; R ^a1 and R ^a2 are each independently H or C _1-6 alkyl; R ⁿ¹ , R ⁿ² and R ⁿ³ are each independently H or C _1-6 alkyl; When Z ² is C _1-6 alkylene, any carbon of the alkylene is optionally substituted with halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, carboxy, C _1-6 alkoxycarbonyl, carbamoyl, C _1-6 alkylcarbamoyl, di(C _1-6 alkyl)carbamoyl, cyano, nitro, oxo or C _6-12 aryl; Z ³ is a direct bond or -C(=O)-; Ring A is a fused benzoheterocyclyl in which a benzene ring is fused to a 6- to 14-membered aryl, a partially unsaturated 9- to 14-membered bicyclic carbocyclyl, a 5- to 12-membered heteroaryl comprising 1 to 3 heteroatoms selected from O, N or S, a 5- to 8-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S, or a 5- to 7-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S; Ring A is optionally substituted with 1 to 3 R ^A ; R ^A is selected from the group consisting of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, cyano, nitro and oxo; Ring E is a 6-14 membered aryl, a partially unsaturated 9-14 membered bicyclic carbocyclyl, a 5-12 membered heteroaryl comprising 1 to 3 heteroatoms selected from O, N or S, a 5-12 membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S, or a fused heterocycloaryl in which a 5-7 membered heterocyclic ring comprising 1 to 3 heteroatoms selected from O, N or S is fused to phenyl; Ring E is optionally substituted with 1 to 3 R ^E ; R ^E is selected from the group consisting of halo; hydroxy; C _1-6 alkoxy; amino; C _1-6 alkylamino; di(C _1-6 alkyl)amino; carboxy; C _1-6 alkoxycarbonyl; carbamoyl; C _1-6 alkylcarbamoyl; di(C _1-6 alkyl)carbamoyl; halosulfonyl; sulfoxy; C _1-6 alkylsulfonyl; cyano; nitro; oxo; or C _1-6 alkyl optionally substituted with one or more of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, nitro, and cyano. In the first paragraph, A compound, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof, selected from compounds represented by the following chemical formula 2 or 3; [Chemical formula 2] [Chemical Formula 3] In the above chemical formulas 2 and 3, Z ¹ , Z ² , X ¹ , X ² , U, R ⁿ¹ , R ⁿ² , ring A and ring E are as described in claim 1. In the second paragraph, A compound, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof, wherein Y ¹ of chemical formula 2 and Y ² of chemical formula 3 are each independently S or O. In the first paragraph, U is selected from NR ⁿ² ; piperidinediyl; piperazinediyl; or a combination of NR ²ⁿ and piperidinediyl or piperazinediyl, a compound, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof. In the first paragraph, U is selected from the group consisting of NR ⁿ² and the following structures, wherein * ¹ of the following structures is connected to a carbon atom of -C(=X ² )-, and * ² is connected to Z ² , a compound, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof: . In the first paragraph, A compound, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof, wherein ring A is phenyl, biphenylyl, tetrahydronaphthyl, pyranyl, pyranonyl, benzopyranyl or benzopyronyl. In the first paragraph, Ring A is or A compound, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof, wherein ring A is optionally substituted with 1 to 3 R ^A . In Article 7, Z ³ is a direct bond, and ring A is , or Either; or Z ³ is -C(=O)-, and ring A is or A compound, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof. In the first paragraph, R ^A is a compound, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof, wherein R A is selected from the group consisting of F, Cl, Br, hydroxy, methoxy, dimethylamino, methyl or oxo. In the first paragraph, Ring A is a compound, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof, selected from the following chemical structures: . In the first paragraph, A compound, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof, wherein ring E is phenyl, naphthalenyl, indanyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, imidazolyl, pyrazolyl, triazolyl, thiophenyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, morpholinyl, thiomorpholinyl, piperidinyl, hexahydroxypyridazinyl, hexahydroxypyrimidinyl, piperazinyl, indolyl, indazolyl, benzoimidazolyl, quinolinyl, isoquinolinyl, cinolinyl, quinazolinyl, quinoxalinyl, phthalazinyl or benzodioxolyl. In the first paragraph, A compound, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof, wherein ring E is selected from the following chemical structures; and ring E is optionally substituted with 1 to 3 R ^E : . In the first paragraph, R ^E is a compound, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof, wherein R E is selected from the group consisting of methoxy, hydroxy, amino, F, Cl, Br, trifluoromethyl, cyano, hydroxymethyl, methyl, fluorosulfonyl, nitro, and oxo. In the first paragraph, Ring E is a compound selected from the following chemical structures, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof: . In the first paragraph, A compound represented by any of the following chemical formulae I to III, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof; [Chemical Formula I] In the above chemical formula I, Y ¹ is S, O or NR ^a1 ; R ^a1 is H or C _1-6 alkyl; R ⁿ¹ and R ⁿ² are each independently H or C _1-6 alkyl; Z ⁴ is a direct bond or C _1-4 alkylene; R ¹ is H, halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, carboxy, C _1-6 alkoxycarbonyl, carbamoyl, C _1-6 alkylcarbamoyl, di(C _1-6 alkyl)carbamoyl, cyano, nitro, oxo or C _6-12 aryl; Ring A is a fused benzoheterocyclyl in which a benzene ring is fused to a 6- to 14-membered aryl, a partially unsaturated 9- to 14-membered bicyclic carbocyclyl, a 6- to 12-membered heteroaryl comprising 1 to 3 heteroatoms selected from O, N or S, a 5- to 8-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S, or a 5- to 7-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S; Ring A is optionally substituted with 1 to 3 R ^A ; R ^A is selected from the group consisting of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, cyano, nitro and oxo; Ring E is a 6-14 membered aryl, a partially unsaturated 9-14 membered bicyclic carbocyclyl, a 5-12 membered heteroaryl comprising 1 to 3 heteroatoms selected from O, N or S, a 5-12 membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S, or a fused heterocycloaryl in which a 5-7 membered heterocyclic ring comprising 1 to 3 heteroatoms selected from O, N or S is fused to phenyl; Ring E is optionally substituted with 1 to 3 R ^E ; R ^E is selected from the group consisting of halo; hydroxy; C _1-6 alkoxy; amino; C _1-6 alkylamino; di(C _1-6 alkyl)amino; carboxy; C _1-6 alkoxycarbonyl; carbamoyl; C _1-6 alkylcarbamoyl; di(C _1-6 alkyl)carbamoyl; halosulfonyl; sulfoxy; C _1-6 alkylsulfonyl; cyano; nitro; oxo; or C _1-6 alkyl optionally substituted with one or more of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, nitro and cyano; [Chemical Formula II] In the above chemical formula II, Y ² is S, O or NR ^a1 ; R ^a1 is H or C _1-6 alkyl; R ⁿ¹ and R ⁿ² are each independently H or C _1-6 alkyl; Z ⁴ is a direct bond or C _1-4 alkylene; R ¹ is H, halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, carboxy, C _1-6 alkoxycarbonyl, carbamoyl, C _1-6 alkylcarbamoyl, di(C _1-6 alkyl)carbamoyl, cyano, nitro, oxo or C _6-12 aryl; Ring A ¹ is a 6- to 14-membered aryl or a partially unsaturated 9- to 14-membered bicyclic carbocyclyl; Ring A ¹ is optionally substituted with 1 to 3 R ^A ; R ^A is selected from the group consisting of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, cyano, nitro and oxo; Ring E is a 6- to 14-membered aryl, a 5- to 12-membered heteroaryl comprising 1 to 3 heteroatoms selected from O, N or S, a 5- to 12-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S, or a fused heterocycloaryl in which a 5- to 7-membered heterocyclic ring comprising 1 to 3 heteroatoms selected from O, N or S is fused to phenyl; Ring E is optionally substituted with 1 to 3 R ^E ; R ^E is selected from the group consisting of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, carboxy, C _1-6 alkoxycarbonyl, carbamoyl, C _1-6 alkylcarbamoyl, di(C _1-6 alkyl)carbamoyl, halosulfonyl, sulfoxy, C _1-6 alkylsulfonyl, cyano, nitro and oxo; [Chemical Formula III] In the above chemical formula III, U ¹ is a direct bond or NR ⁿ² ; At least one of Y ³ and Y ⁴ is N, and the others are CH; Z ⁵ is a direct bond, C _1-6 alkylene, -NR ⁿ³ CO- or a 5- to 7-membered heteroaryl comprising 1 to 3 heteroatoms selected from N, O and S; R ⁿ¹ , R ⁿ² and R ⁿ³ are each independently H or C _1-6 alkyl; Ring A is a fused benzoheterocyclyl in which a benzene ring is fused to a 6- to 14-membered aryl, a partially unsaturated 9- to 14-membered bicyclic carbocyclyl, a 5- to 12-membered heteroaryl comprising 1 to 3 heteroatoms selected from O, N or S, a 5- to 8-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S, or a 5- to 7-membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S; Ring A is optionally substituted with 1 to 3 R ^A ; R ^A is selected from the group consisting of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, cyano, nitro and oxo; Ring E is a 6-14 membered aryl, a partially unsaturated 9-14 membered bicyclic carbocyclyl, a 5-12 membered heteroaryl comprising 1 to 3 heteroatoms selected from O, N or S, a 5-12 membered heterocyclyl comprising 1 to 3 heteroatoms selected from O, N or S, or a fused heterocycloaryl in which a 5-7 membered heterocyclic ring comprising 1 to 3 heteroatoms selected from O, N or S is fused to phenyl; Ring E is optionally substituted with 1 to 3 R ^E ; R ^E is selected from the group consisting of halo; hydroxy; C _1-6 alkoxy; amino; C _1-6 alkylamino; di(C _1-6 alkyl)amino; carboxy; C _1-6 alkoxycarbonyl; carbamoyl; C _1-6 alkylcarbamoyl; di(C _1-6 alkyl)carbamoyl; halosulfonyl; sulfoxy; C _1-6 alkylsulfonyl; cyano; nitro; oxo; or C _1-6 alkyl optionally substituted with one or more of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, nitro and cyano; s is 0 or 1. In Article 15, A compound represented by Formula IA, Formula IB, Formula IC, Formula ID, Formula IE or Formula IIA, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof; [Chemical Formula IA] [Chemical Formula IB] [Chemical Formula IC] [chemical formula ID] [chemical formula IE] [Chemical formula IIA] In the above chemical formulas IA, IB, IC, ID, IE and IIA, R ^A is selected from the group consisting of halo, hydroxy, C _1-6 alkoxy, amino, C _1-6 alkylamino, di(C _1-6 alkyl)amino, C _1-6 alkyl, cyano, nitro and oxo; n, m1, m2, o1, o2, p and q are each independently an integer from 0 to 3; The sum of m1 and m2 and o1 and o2 is each less than or equal to 3; Y ¹ , Y ² , R ⁿ¹ , R ⁿ² , Z ⁴ , R ¹ and ring E are as described in claim 13. In the first paragraph, A compound, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof, selected from compounds represented by the following chemical formula: . A pharmaceutical composition for preventing or treating a disease associated with decreased expression of SOX9 (SRY-Box Transcription Factor 9), comprising a compound of any one of claims 1 to 17, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof. In Article 18, A pharmaceutical composition, wherein the disease associated with the decreased expression of the above SOX9 is osteoarthritis. A method for treating a disease associated with decreased expression of SOX9, comprising administering to a subject a compound of any one of claims 1 to 17, a stereoisomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof. Use of a compound according to any one of claims 1 to 17, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof, for the prevention or treatment of a disease associated with decreased expression of SOX9. Use of a compound according to any one of claims 1 to 17, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the prevention or treatment of a disease associated with decreased expression of SOX9.